Objective:To develop a simple and efficient method for constructing a middle fragment-deleted mutant of MHC class Ⅱ molecule transactivator(CⅡTA)mutant with the 109~(th)to 226~(th) amino acid codons deleted.Methods: Two gene fragments at each end of the deleted CⅡTA gene were obtained by OE-PCR method and were mixed together for 8 PCR cycles without primers to achieve effective overlapping,then 2 primers was added for amplification of the desired fragments.The amplification products were subsequently cloned into eukaryotic vector pIRES for identification.Results: A mutant of CⅡTA with the 109~(th)to 226~(th) amino acids deleted was successfully constructed.Conclusion: This modified OE-PCR technique overcomes some shortcomings of traditional method and is very suitable for constructing mutants with middle fragment deletion,making it worth to be popularized.

Objectives:To understand the human resources and costs of essential public health services based on community health service centers in Beijing’s rural areas and to offer policy recommendations for advancing the progress of essential public health services. Methods:With the self-designed questionnaire, the paper collected data from the districts of Changping, Tongzhou, Huairou and Miyun. By cluster random sampling, 12 community health service centers and 108 personnel were surveyed based on the level of economic development. Results: The number of young women, preventive personnel and staff with low ranking positions represents a significant share of all essen-tial public health service providers. Beijing’s rural areas deploy a great deal of human resources for vaccination and health education with little set aside for women’s health and reproductive health. The unit cost of setting special col-umns for health education and field disease treatment is higher than other service items. The suburbs deploy a great deal of human resources for service items than the outskirts while the unit costs are lower. Conclusions: Human re-source for essential public health services in Beijing’s rural areas should be strengthened. The costs of service items differ greatly so that policy adjustments and finances should be implemented based on actual demand and resources.

Objective To investigate the leptin in continuous ambulatory peritoneal dialysis(CAPD)and its influen- cing factors.Methods 30 CAPD patients and 30 henhhy controls were enrolled in the study.Leptin was measured by ELISA.C reactive protein(CRP)by transmisson turbidmetry.Body mass index(BMI),fasting insulin(FINS), blood urea nitrogen(BUN),serum creatinine(Scr)etc were also determined.These patients were classified to two nutritional categories based on SGA normal nutrition and malnutrition.The relationship between serum leptin concentra- tion and these indices was analysed.Results Serum leptin levels in CAPD group was significantly higher than those of healthy control(P

Epidemiological studies have shown that patients with diabetes are prone to have depression and anxiety.This article reviews the evidence,mechanism,complications and strategy of selective serotonin reuptake inhibitors paroxetine in treatment of depression and anxiety in patients with type 2 diabetes mellitus.

Objective: To develop a simple and efficient method for constructing a middle fragment-deleted mutant of MHC class II molecule transactivator (C II TA) mutant with the 109th to 226th amino acid codons deleted. Methods: Two gene fragments at each end of the deleted C II TA gene were obtained by OE-PCR method and were mixed together for 8 PCR cycles without primers to achieve effective overlapping, then 2 primers was added for amplification of the desired fragments. The amplification products were subsequently cloned into eukaryotic vector pIRES for identification. Results: A mutant of C II TA with the 109th to 226th amino acids deleted was successfully constructed. Conclusion: This modified OE-PCR technique overcomes some shortcomings of traditional method and is very suitable for constructing mutants with middle fragment deletion, making it worth to be popularized.

ObjectiveTo study the differential expression of signal transduction related gene between the tunica media in human abdominal aortic aneurysms (AAA) and normal aorta, and inspect their role in the etiology of abdominal aortic aneurysms.MethodsGene chip technique was applied to study the differential expression of signal transduction related gene in human AAA, then screening some significant differential expressed genes for further research by RT-PCR, Western Blot, and immunohistochemistry.ResultsForty-five differentially expressed genes related to cellular signal transduction were detected accounting for 1.09% of the chip′s total 4?096 genes,among them 28 genes were up-regulated and 17 down-regulated in aortic aneurysm. ASK1, ERK1 gene were found up-requlated in aortic aneurysm.ConclusionsDifferential expression of signal transduction gene could play a key role in the occurrence of AAA.

Objective To investigate the common causes of acute renal failure(ARF) in the elderly hospitalized patients and the effect of continuous peritoneal dialysis(PD) by bedside intubation.Methods The data of hospitalized patients with ARF including 28 elderly patients (mean age 68.2?9.6 yrs) and 13 adult patients (mean age 39.4?9.1 yrs) were studied.Of them,32 patients received PD by bedside intubation.The nosogenesis of ARF was analyzed and the effect of emergency PD by bedside cmpared between two groups.Results Among various causes,prerenal factors accounted for 32.1%,infection and wound of bad control 39.3%,unreasonable use of nephrotoxic drugs 14.3%,which totally accounted for 85.7%.Of 21 elderly patients (76%) receiving PD treatment,11 (52.4%) acquired completely remission,5 (23.8%) turned to chronic PD,the successful rate of intensive care being 81.0%,and the death rate being 19.1%.Conclusion Prerenal factors,infection,wound of bad and unreasonable use of nephrotoxic drugs are the common causes of ARF in the elderly hospitalized patients.PD by bedside intubation is safe,simple and effective.

Objective To approach the significance of serum protein fingerprinting on the identification of different courses of acute myocardial infarction. Methods For 153 patients of acute myocardial infarction, including 45 cases of just admitted patients, 12 cases of 3h admitted patients, 22 cases of 6h admitted patients, 24 cases of 9h admitted patients, 24 cases of 12h admitted patients, 16 cases of 24h admitted patients and 10 cases of 48h admitted patients. The relative contents of serum proteins were detected by IMAC3 chip and proteinchip reader (CipherGen Inc., VS). Results On the M/Z values ranged from 4KDa to 12KDa, ten protein contents were obviously different between the different courses of acute myocardial infarction. All the patients in different courses were diagnosed correctly, the accuracy was 100%(153/153). Both sensitivity and specificity were also 100% in learning mode. Conclusion Different courses of acute myocardial infarction patients can be quickly and correctly diagnosed by this method with high sensitivity and specificity. That will be widely used in clinical application

Acid-sensing ion channels( ASICs )are ubiquitously expressed both in periphery nervous system, where they are involved in nociception,mechanosensation,inflammation ,cardiac angina, and in central nervous system,where they are essential to a variety of physiologic and pathophysiologic processes,such as synaptic plasticity ,learning ,memory and ischemic brain injury. Here in the article, we present a collection of key points about ASICs, ranging from molecular identity and expression, regulatory mechanisms responsible for channel gating to their multiple functions. Finally, a future prospect for the investigation of ASICs is outlined.

Objective:To construct adenovirus containing murine MHC class II transactivator (CIITA) mutant gene and to observe expression of this gene mediated by adenovirus as well as function of the expression product in vitro.Methods:The type IV CIITAcDNA gene was cloned from peritoneal macrophage of BALB/c mouse by conventional way of molecular biology, and then it was cloned into expression vector pIRES. CIITA mutant gene was constructed by the way of overlap extension by PCR (OE-PCR), and then it was also cloned into expression vector pIRES. By using of pAdEasy-1 system we acquired the deficient recombination adenovirus Ad-CIITAm, containing CIITA mutant gene, which had the ability of infection and the no-loaded control adenovirus Ad-GFP.Then the two adenoviruses strains were processed for a great deal amplification,purification and titer determining. The Ad-CIITAm and Ad-GFP were infected into HeLa cells and Raji cells.Inducible or constitutive expression of HLA-DR molecule and the following changes were observed by the use of flow cytometry.Results:The murine CIITA gene was cloned successfully and the recombinantant adenovirus Ad-CIITAm containing murine CIITA mutant gene was also constructed successfully. It was proved by flow cytometry that expression of HLA-DR molecules on the surface of HeLa cells and Raji cells infected by Ad-CIITAm were all decreased remarkably in comparison with those of control cells infected by Ad-GFP.Conclusion:This experiment proves that expressed murine CIITA mutant mediated by adenovirus could inhibit the expression of MHC II molecules efficiently.